Specific salt, anhydrous and crystalline form of a dihydropteridione derivative

ABSTRACT

The present invention relates to a specific salt of a dihydropteridione derivative, namely the trihydrochloride salt of the compound N-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide, to its crystallisation in the form of an hydrate with water, to a process for the manufacture thereof, and to the use thereof in a pharmaceutical composition.

This application is a continuation of application Ser. No. 11/668,681filed Jan. 30, 2007.

The present invention relates to a specific salt of a dihydropteridionederivative, namely the trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,to its anhydrous form, to its crystallisation in the form of an hydratewith water, to a process for the manufacture thereof, and to the usethereof in a pharmaceutical composition.

BACKGROUND TO THE INVENTION

A number of dihydropteridione derivatives are already known in the priorart. Thus, for example, International Patent Applications WO 03/020722and WO 2004/076454 disclose dihydropteridione derivatives, a process fortheir manufacture and their use in a pharmaceutical composition to treatdiseases connected with the activity of specific cell cycle kinases andcharacterised by excessive or abnormal cell proliferation.

The compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideand a process for its manufacturing are specifically disclosed in WO2004/076454.

However, the above-mentioned patent applications do not disclose anyspecific salt form or crystal form of any of the compounds exemplifiedtherein.

Although the pharmacologically valuable properties of thedihydropteridinone derivatives disclosed in the art and mentioned aboveconstitute the basic prerequisite for effective use of the compounds aspharmaceutical compositions, an active substance must in any casesatisfy additional requirements in order to be accepted for use as adrug. These parameters are largely connected with the physicochemicalnature of the active substance. Hence, there continues to be a need fornovel salt and crystalline forms of active substances, which can beconveniently formulated for administration to patients and which arepure and highly crystalline in order to fulfil exacting pharmaceuticalrequirements and specifications.

Preferably, such compounds will be readily formed and have favourablebulk characteristics. Examples of favourable bulk characteristics aredrying times, filterability, solubility, intrinsic dissolution rate,stability in general and especially thermal stability, andhygroscopicity.

An absence of breakdown products in the pharmaceutical composition beingused is also favourable, since if breakdown products are present in thepharmaceutical composition the content of active substance present inthe pharmaceutical formulation might be lower than specified.

Another critical parameter to be controlled is the hygroscopicity, sincethe absorption of moisture reduces the content of pharmaceuticallyactive substance as a result of the increased weight caused by theuptake of water. Pharmaceutical compositions with a tendency to absorbmoisture have to be protected from moisture during storage, e.g. by theaddition of suitable drying agents or by storing the drug in anenvironment where it is protected from moisture. In addition, the uptakeof moisture may reduce the content of pharmaceutically active substanceduring manufacture if the pharmaceutical substance is exposed to theenvironment without being protected from moisture in any way.Preferably, therefore, the hygroscopicity of a pharmaceutically activesubstance should be well characterised, and possibly also stabilized.

As the crystal modification of an active substance is important to thereproducible active substance content of a preparation, there is a needto clarify as far as possible any existing polymorphism of an activesubstance present in crystalline form. If there are differentpolymorphic modifications of an active substance care must be taken toensure that the crystalline modification of the substance does notchange in the pharmaceutical preparation later produced from it.Otherwise, this could have a harmful effect on the reproducible potencyof the drug. Against this background, active substances characterised byonly slight polymorphism are preferred.

Decreased levels of organic solvents in the crystal lattice are alsofavourable, due in part to potential solvent toxicity to the recipientas a function of the solvent.

Another criterion which may be of exceptional importance under certaincircumstances depending on the choice of formulation or the choice ofmanufacturing process is the solubility of the active substance. If forexample pharmaceutical solutions are prepared (e.g. for infusions) it isessential that the active substance should be sufficiently soluble inphysiologically acceptable solvents. For drugs which are to be takenorally, it is in general very important that the active substance shouldbe sufficiently soluble and bioavailable.

Furthermore, the process for preparing such a compound also needs to beconveniently carried out on commercial scale.

Hence, without being restrictive, examples of the parameters which needsto be controlled are the stability of the starting substance undervarious environmental conditions, the stability during production of thepharmaceutical formulation and the stability in the final compositionsof the drug.

The pharmaceutically active substance used to prepare the pharmaceuticalcompositions should therefore have great stability which is ensured evenunder all kinds of environmental conditions.

The problem of the present invention is thus to provide apharmaceutically active substance which is not only characterised byhigh pharmacological potency but also satisfies the above-mentionedphysicochemical requirements as far as possible.

SUMMARY OF THE INVENTION

Surprisingly, it has been found that the problem outlined above issolved by the trihydrochloride salt and by the crystallinetrihydrochloride hydrate form of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,the structure of which is depicted below in the form of the free base asformula (I).

The trihydrochloride salt and the crystalline trihydrochloride hydrateform of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideoffer numerous advantages over the free base and the anhydrous form ofthis compound. Hence, over the anhydrous form, the crystallinetrihydrochloride hydrate form presents for example an advantage ofstability under ambient conditions.

Furthermore, when compared to the base salt, the trihydrochloride saltpresents an advantage of solubility in water, where the trihydrochlorideis much more soluble than the free base.

Lastly, when compared to other common salt form builders, thetrihydrochloride salt form presents also advantages as far as the yield,purity and reproducibility of the synthesis of the salt form areconcerned.

Thus, a first object of the present invention is the trihydrochloridesalt form of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.

A further object of the present invention is the trihydrochlorideanhydrous form of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.

A further object of the present invention are the crystallinetrihydrochloride hydrate forms of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6oxo-2-pteridinyl]amino]-3-methoxy-benzamide.

A further object of the present invention is the crystallinetrihydrochloride trihydrate form of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.

A further object of the present invention is an improved process for themanufacturing of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.

A further object of the present invention is a process for themanufacturing of the trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.

A further object of the present invention is a pharmaceuticalcomposition comprising the trihydrochloride salt, a crystallinetrihydrochloride hydrate form or the crystalline trihydrochloridetrihydrate form of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,together with one or more pharmaceutically acceptable carriers, diluentsor excipients.

A further object of the present invention is the use of thetrihydrochloride salt, a crystalline trihydrochloride hydrate form orthe crystalline trihydrochloride trihydrate form of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,in a method for treating diseases characterised by excessive or abnormalcell proliferation, or for preparing a pharmaceutical composition whichis suitable for treating diseases characterised by excessive or abnormalcell proliferation.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the X-ray powder diffractogram of the crystallinetrihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,recorded using a Bruker D8 Advanced-diffractometer fitted with alocation-sensitive detector (OED) and a Cu anode as the x-ray source(CuKα radiation, λ=1.54056 Å, 40 kV, 40 mA).

FIG. 2 shows a light microscopy photograph of crystals of thetrihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.

FIG. 3 shows the thermoanalysis and determination of the melting point(DSC/TG) of the trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,recorded using a DSC and evaluated by the peak onset (heating rate: 10°C./min). The value given is determined using a DSC 821e made by MettlerToledo.

DETAILED DESCRIPTION OF THE INVENTION

As already mentioned hereinbefore, the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideis specifically disclosed in WO 2004/076454, as well as a process forits preparation. For details on a process to manufacture this compound,reference is thus made to this patent application.

For an alternative process to manufacture this compound, reference ismade to the following process.

Abbreviations used.

-   -   TLC Thin-Layer Chromatography    -   DSC Differential Scanning Calorimeter    -   TG ThermoGravimetry

The starting materials trans-4-aminocyclohexanol 10,3-methoxy-4-nitrobenzoic acid 2, N-(cyclopropylmethyl)piperazine 12c,and 4-acetamido-cyclohexanone 18 are known compounds which arecommercially available.

This process is a convergent process, which includes the steps of:

(i) synthesis of a compound of formula 15c

(ii) synthesis of a compound of formula 16

and (iii) reacting the compound of formula 15c with the compound offormula 16.

-   -   In the foregoing, the synthesis of a compound of formula 15c and        the synthesis of a compound of formula 16 are described.    -   (i) Synthesis of a compound of formula 15c

260 g (1.32 mol) 3-methoxy-4-nitrobenzoic acid 2 are placed in 1.5 Ltoluene. 300 mL toluene are distilled off. 5 mL dimethylformamide areadded to the residue and 123 mL (1.7 mol) thionyl chloride are addeddropwise thereto. The reaction solution is refluxed for 2 hours. Thesolvent is concentrated by evaporation using the rotary evaporator underreduced pressure. The residue is dissolved in 500 mL tetrahydrofuran andadded dropwise to a suspension of 202 g (1.33 mol)trans-4-aminocyclohexanol 10 in 1.5 L tetrahydrofuran and 1.38 L of a30% potassium carbonate solution, so that the temperature is maintainedbetween 5° and 13° C. The mixture is stirred for 1 hour at 20° C. and 5L demineralised water are added. The precipitate is suction filtered andwashed with demineralised water. The solid is dried at 70° C. in thecirculating air dryer. 380 g (98% of theory) product 11 are obtained.TLC (methylene chloride/ethanol=9:1) R_(f)=0.47

1 g of finely powdered ruthenium(III)-chloride hydrate are added to 185g (0.63 mol) 11 and 234 g N-methylmorpholine-N-oxide in 1.8 Lacetonitrile and the mixture is refluxed for 1 hour. Under reducedpressure 1.6 L acetonitrile are evaporated off. 1.5 L demineralisedwater are added to the residue and the suspension is cooled to 5° C. Theprecipitate is suction filtered and washed with plenty of demineralisedwater. The solid is dried at 70° C. in the circulating air dryer. 168 g(91% of theory) product 13 are obtained.

TLC (methylene chloride/ethanol=9:1) R_(f)=0.64

112 g (383 mmol) of product 13, 108 g (770 mmol)N-(cyclopropylmethyl)piperazine 12c and 4.5 mL methanesulphonic acid intoluene are refluxed for 3 hours using the water separator (approx. 76mL water are separated off). Under reduced pressure 900 mL toluene areevaporated off and the residue is suspended in 1.2 L ethanol. 15 gsodium borohydride are added batchwise to this suspension at atemperature of 15° to 25° C. within one hour. The mixture is stirred for3 hours at 20° C. and another 4 g sodium borohydride are added. Themixture is stirred for 16 hours at 20° C. Under reduced pressure 650 mLethanol are evaporated off. 2 L demineralised water and 300 mLcyclohexane are added. The mixture is cooled to 5° C. and the suspensionis suction filtered. The residue is dissolved in 1 normal hydrochloricacid. 5 g activated charcoal are added and the mixture is suctionfiltered. 400 mL tert.-butylmethylether are added to the filtrate and itis made alkaline with ammonia solution. It is cooled to 4° C., theprecipitate is suction filtered and washed with demineralised water. Theresidue is refluxed in 400 mL tert.-butylmethylether. It is cooled, thesolid is suction filtered and washed with tert.-butylmethylether. Afterdrying in the circulating air dryer at 60° C. 73 g (46% of theory)product 14c is obtained.

TLC (methylene chloride/ethanol=9:1) R_(f)=0.2

The compound J may alternatively also be prepared by the followingmethod.

22 g (142 mmol) 4-acetamido-cyclohexanone 18, 39.7 g (283 mmol)N-cyclopropylmethylpiperazine 12c and 0.71 mL methanesulphonic acid in175 mL toluene are refluxed using the water separator until no morewater is precipitated. The mixture is left to cool and at 50° C. 175 mLethanol are added and the resulting mixture is cooled to 20° C. 5.37 g(142 mmol) sodium borohydride are added batchwise with thorough stirringand the mixture is stirred for 16 hours at 20° C. 200 mL of 4 normalhydrochloric acid are added dropwise to the reaction mixture. Underreduced pressure 200 mL solvent are evaporated off. 100 mL saturatedpotassium carbonate solution and 200 mL methylisobutylketone are addedto the residue. The two-phase mixture is cooled to 5° C. with thoroughstirring. The product is suction filtered and dissolved at refluxtemperature in 90 mL methylisobutylketone. After the addition ofactivated charcoal it is filtered hot. The mixture is left to cool andthe precipitate is removed by suction filtration. After drying, 16.2 g(41% of theory) of trans compound 19 are obtained.

TLC (methylene chloride/ethanol/ammonia=9:1:0.1) R_(f)=0.39

A solution of 44 g (157 mmol) of product 19 in 500 mL 24% hydrochloricacid is refluxed for 6 hours. The solvent is concentrated by evaporationunder reduced pressure and the residue is crystallised from 700 mLisopropanol. The precipitate is suction filtered, washed withtert.-butylmethylether and dried at 60° C. in the vacuum dryingcupboard. 54.7 g product 20 are obtained as the trihydrochloride(contains 5% water).

33 g (90.4 mmol) 3-methoxy-4-nitrobenzoic acid 2 are suspended in 80 mLtoluene. 0.5 mL dimethylformamide and 16 g (134 mmol) thionyl chlorideare added. The mixture is refluxed for 1 hour. The solution isconcentrated by evaporation under reduced pressure and the crude acidchloride is dissolved in 50 mL tetrahydrofurane. The solution is addeddropwise to a suspension of 18.7 g (94.9 mmol, 95%) of 20trihydrochloride and 49 g (397 mmol) of diisopropylethylamine in 150 mLtetrahydrofurane while being cooled in the ice bath. TLC is used tocheck that the reaction is complete. After the reaction has ended wateris added to the suspension and the pH is adjusted to 10 by the additionof sodium hydroxide solution. The organic phase is separated off andwashed with saturated saline solution. The combined aqueous phases areextracted once with tetrahydrofurane. The combined organic phases areconcentrated by evaporation under reduced pressure. The residue isrefluxed in 300 mL tert.-butylmethylether. The mixture is left to coolto 20° C. and the precipitate is suction filtered. After drying in thevacuum drying cupboard at 45° C., 31.3 g (83% of theory) of product 14cis obtained.

A solution of 72.5 g (174 mmol) of product 14c in 700 mL methanol and145 mL dimethylformamide is hydrogenated in the presence of 10 g Raneynickel at a temperature of 20° C. and a hydrogen pressure of 50 psi. Thecatalyst is filtered off and the methanol is evaporated under reducedpressure. 500 mL demineralised water are added to the residue and thesuspension is cooled to 5° C. The precipitate is suction filtered andwashed with demineralised water. After drying in the circulating airdryer at 60° C., 60.5 g (90% of theory) of product 15c is obtained.

TLC (methylene chloride/ethanol/ammonia=9:1:0.1) R_(f)=0.58

-   -   (ii) Synthesis of a compound of formula 16

The synthesis of the2-chloro-7-ethyl-7,8-dihydro-5-methyl-8-(1-methylethyl)-(7R)-6(5H)-pteridinone16 is described in general in WO 2004/076454, to which reference ismade.

The present invention provides an alternative route of synthesis of the2-chloro-7-ethyl-7,8-dihydro-5-methyl-8-(1-methylethyl)-(7R)-6(5H)-pteridinone16, which is described in the following.

The following starting materials are known and commercially available:(R)-2-amino-butyric acid 21 and 2,4-dichloro-5-nitropyrimidine 5.

A suspension of 25 g (242 mmol) (R)-2-amino-butyric acid 21 and 32 mL(290 mmol) trimethylorthoformate in 150 mL methanol is heated to 50° C.At this temperature 26.5 mL (364 mmol) of thionylchloride are added in30 minutes. Under evolution of gas the temperature increases to 60° C.The reaction mixture is refluxed for 3 hours. 125 mL methanol aredistilled off and 100 mL toluene are added. 75 mL of solvent are removedby distillation. A suspension of 77 g (364 mmol) sodiumtriacetoxyborohydride in 175 mL toluene is added to the reaction mixtureat 60° C. 22 mL acetone are added at 40° C. The reaction mixture isstirred for 16 hours at room temperature. Under cooling 73 mL ammonia(25%) is added. After addition of 50 mL of demineralised water themixture is heated to 50° C. The organic phase is separated and washedwith demineralised water. 24 mL of a 10 molar solution ofhydrogenchloride in ethanol is added. 125 mL of solvent are removed bydistillation. 175 mL tetrahydrofurane is added and the suspension iscooled to 2° C. The suspension is suction filtered and washed with coldtetrahydrofurane. After drying in a vacuum drying oven at 50° C., 42.9 g(90% of theory) of product 22 as hydrochloride is obtained.

9.3 mL of a 50% aqueous sodium hydroxide solution is added to a stirredmixture of 33.3 g (170 mmol) 22 hydrochloride in 60 mL cyclohexane and60 mL demineralised water. The aqueous phase is separated and theorganic phase is added dropwise to a refluxed suspension of 30 g (155mmol) 5 and 52 g (619 mmol) sodium hydrogencarbonate in 230 mLcyclohexane. The suspension is refluxed for 5 hours using a waterseparator to remove the formed water. 75 mL of solvent is destined off.At 75° C. the suspension is suction filtered to remove the salts. Thesolvent is destined of. The residue is dissolved in 240 mL 2-propanoland 90 mL of solvent is destined of again. The solution is cooled slowlyto 2° C. The suspension is suction filtered and washed with cold2-propanol. After drying in a vacuum drying oven at 50° C., 38.9 g (79%of theory) of product 23 is obtained

Lipophilic solvents such as e.g. cyclohexane, methylcyclohexane, tolueneand the mixtures thereof are particularly suitable for achieving highregioselectivity in the nucleophilic substitution reaction with compound5.

50 g 23 in 375 mL of tetrahydrofurane is hydrogenated in the presence of5 g Platinum on Carbon (5%) at a hydrogene pressure of 3 bar and at 35°C. until no further hydrogene consumed. 2.5 g vanadyl acetylacetonateare added and the hydrogenation is continued. The suspension is filteredto remove the catalysts. The solvent is removed under reduced pressure.150 mL 2-propanol are added to the residue and heated to reflux. 300 mlof demineralised water are added. The suspension is cooled slowly to 2°C. The suspension is suction filtered and washed with a cold mixture of2-propanol and demineralised water. After drying in a vacuum drying ovenat 50° C., 36 g (90% of theory) of product 24 is obtained

A suspension of 7 g (27.5 mmol) of product 25 and 5.7 g (41 mmol)potassium carbonate in 30 mL dimethylcarbonate is heated to 130° C. inan autoclave for 5 hours. The mixture is left to cool and 25 mLdemineralised water and 15 mL ethyl acetate are added with stirring. Theorganic phase is distilled off under reduced pressure. A mixture of 25mL ethanol and 45 mL demineralised water is added to the residue andheated to 60° C. The solution is left to cool to room temperature. Theprecipitate is suction filtered and washed with a mixture ofdemineralised water and ethanol (2:1). The product is dried at 50° C. inthe vacuum drying cupboard. 6 g (82% of theory) of product 16 areobtained.

-   -   (iii) Reaction of a compound of formula 15c with a compound of        formula 16

A solution of 23 g (59.5 mmol) of compound 15c, 16.8 g (62.5 mmol)2-chloro-7-ethyl-7,8-dihydro-5-methyl-8-(1-methylethyl)-(7R)-6(5H)-pteridinone16 and 28.3 g (149 mmol) para-toluenesulphonic acid hydrate in 350 mL2-methyl-4-pentanol is refluxed for 22 hours using the water separator.After the addition of 1 g of compound 16 the mixture is refluxed for afurther 2 hours. 300 mL solvent are distilled off and the viscous oil isallowed to cool to 60° C. 300 mL methylene chloride and 300 mLdemineralised water are added and the pH is raised by adding approx. 20mL of 10 normal sodium hydroxide solution to pH=9. The organic phase iswashed twice with demineralised water and dried over sodium sulphate.The solvent is evaporated off under reduced pressure and the residue isdissolved at 65° C. in 200 mL ethyl acetate. The mixture is left to coolslowly to 20° C., the precipitate is suction filtered and washed withcold ethyl acetate. After drying at 60° C. in the vacuum drying cupboard24.4 g (66% of theory) of product (I) is obtained (m.p.=182° C., DSC: 10K/min, additional endothermic effects in the DSC diagram beforemelting).

Thus, it is an object of the present invention to provide a process forthe manufacture of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I)

-   -   characterised in that a compound of formula 15c

-   -   is reacted with a compound of formula 16,

-   -   wherein the compound of formula 16 is prepared by methylation of        a compound of formula 8

-   -   in the presence of dimethylcarbonate.

The present invention further provides a process for the manufacture ofthe trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I), said process comprising the steps of contacting, underelevated temperature or at room temperature,N-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamidedissolved in a suitable solvent or in a mixture of solvents, withhydrochloric acid or hydrogen chloride gas dissolved in an organicsolvent, optionally in the presence of para-toluenesulfonic acid, andcollecting the precipitate formed. Suitable solvents to dissolveN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamidefor performing the salt formation are alcohols like methanol, ethanol,1- or 2-propanol, isomeric alcohols of butanol, isomeric alcohols ofpentanol, isomeric alcohols of hexanol, like 2-methyl-4-pentanol,ketones like acetone, dialkylethers like tetrahydrofurane, acetic acidesters like ethyl acetate, organic acids like acetic acid, amides likeN-methylpyrrolidinone and nitrites like acetonitrile.

An alternative manufacturing process is illustrated by the followingexperiment, in which the trihydrochloride salt is obtained by additionof concentrated hydrochloric acid to the reaction medium aftercompletion of the acid mediated nucleophilic aromatic substitutionreaction of the compound 15c with the compound 16. The following exampleis illustrative of the present invention and therefore is not to beregarded as a limitation to its scope.

-   -   A suspension of 143 g (0.37 mol) 15c and 110 g (0.41 mol) 16 in        2 L 2-methyl-4-pentanol is heated up to 60° C. 176 g (0.93 mol)        para-toluenesulfonic acid monohydrate are added and the mixture        is heated to reflux for 24 hours using a water separator. The        solution is cooled to 100 ° C. 183 g concentrated hydrochloric        acid are added. At 60° C. 1.5 L acetone are added. The        suspension is stirred for 16 hours at room temperature. The        precipitate is suction filtered and washed with acetone. The        product is dried at 60° C. in the vacuum drying cupboard. 267 g        (92% of theory) of compound (I) as trihydrochloride are        obtained.

A further object of the present invention is the following step ofpurification of the trihydrochloride salt via crystallization, wherein:

-   -   the compound (I) as trihydrochloride is suspended in a suitable        organic solvent, such as ethanol;    -   the reaction medium is heated to reflux;    -   water is added;    -   after cooling the precipitate is collected, washed with a        suitable solvent, such as ethanol, and dried.

The following example of purification via crystallization isillustrative of the present invention and therefore is not to beregarded as a limitation to its scope.

-   -   Example of purification of the trihydrochloride salt of the        compound of formula (I) via crystallization.

A suspension of 15.5 g of compound (I) as trihydrochloride in 160 mL dryethanol is heated to reflux. 5.5 mL of demineralised water are added.The solution is left to cool slowly to 20° C. and stirred 16 hours at20° C. The precipitate is suction filtered and washed with ethanol.After drying at 50° C. in the vacuum drying cupboard 13.3 g (86% oftheory) of compound (I) as trihydrochloride is obtained.

A further object of the present invention is a process for themanufacture of an hydrated crystal form of the trihydrochloride salt ofthe compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I), said process comprising the steps of:

-   -   dissolving the compound (I) as base in a suitable organic        solvent, such as ethanol, at room temperature or elevated        temperature;    -   adding hydrochloric acid to the reaction medium;    -   cooling the reaction medium;    -   collecting the precipitate, washing the precipitate with e.g.        ethanol, and drying.

The following example of manufacture of an hydrated crystal form of thetrihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-²-pteridinyl]amino]-3-methoxy-benzamideof formula (I) is illustrative of the present invention and therefore isnot to be regarded as a limitation to its scope.

-   -   Example of manufacture of an hydrated crystal form of the        trihydrochloride salt of the compound        N-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide        of formula (I).

1.1 g of concentrated hydrochloric acid are added to a solution of 2 gof the free base of (I) in 30 mL ethanol. After stirring for 2 hours at20° C. the suspension is cooled to 2° C. The precipitate is suctionfiltered and washed with ethanol. After drying in the vacuum dryingcupboard 2.15 g (91% of theory) of product (I) as trihydrochloride areobtained.

The trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-²-pteridinyl]amino]-3-methoxy-benzamideof formula (I), is represented by the following Formula.

The present invention also provides a process for the manufacture of ananhydrous form of the trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I), wherein a preparation of the trihydrochloride salt ofthe compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I) is dried at a temperature above 130° C. and maintainedunder dry atmosphere.

The following solubility and solid state characteristics of an hydratedform of the trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I), are relevant to the present invention.

-   -   Solubility properties of an hydrated form of the        trihydrochloride salt of        N-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(        1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.        -   Solubility in Aqueous Media

Table I shows the values of solubility of an hydrated form of thetrihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I) in different aqueous media.

TABLE I pH of the Solubility saturated Medium [mg/ml] solution water >103.7 0.1 N HCl >10 1.3 0.01 N HCl >10 n.d. 0.1 mol/l citric acid >10 n.d.0.1 mol/l tartaric acid >10 n.d. 0.01 mol/l >10 n.d. methanesulfonicacid McIlvaine buffer pH 2.2 >10 2.3 McIlvaine buffer pH 3.0 6.2 3.0McIlvaine buffer pH 4.0 7.2 3.9 McIlvaine buffer pH 5.0 7.2 4.7McIlvaine buffer pH 6.0 7.1 5.7 McIlvaine buffer pH 7.4 7.8 7.1 0.1mol/l meglumine 0.017 pH 8.9 0.01 N NaOH >10 pH 7.4 n.d. = notdetermined

From the above results, it can be concluded that this hydrated form ofthe trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I) has a pH dependent solubility profile in aqueous mediawith high solubility in acidic and neutral media and strongly reducedsolubility in basic media due to the lower solubility of the free base.

-   -   Solubility in Organic Media

This hydrated form of the trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I) is highly soluble (>10 mg/ml) in propylenglycol,glycofurol, Cremophor RH40 (30% aqueous solution), Poloxamer 188 (20%aqueous solution), Solutol HS 15 (20% aqueous solution) as well asHP-β-cyclodextrin (20% -, 10% -, 5% - aqueous solutions).

-   -   Solid state properties of an hydrated form of the        trihydrochloride salt of        N-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.        -   Appearance

In the solid state, this hydrated form of the trihydrochloride salt ofthe compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I) appears as a white to off-white microcrystalline powder.

-   -   Crystallinity and Polymorphism

This hydrated form of the trihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I) is highly crystalline. The X-ray powder diffractiondiagram is shown in FIG. 1.

The X-ray powder reflection and intensities (standardised) are shown inthe following Table II.

TABLE II 2 Θ d_(hkl) Intensity [°] [Å] I/I_(o) [%] 4.13 21.38 24 7.8811.21 89 8.25 10.71 34 8.66 10.21 28 9.81 9.01 100 11.45 7.73 54 12.387.14 59 13.12 6.74 37 13.50 6.55 52 14.53 6.09 39 15.26 5.80 32 15.405.75 32 15.51 5.71 37 16.52 5.36 43 16.71 5.30 28 17.35 5.11 10 18.044.91 43 18.19 4.87 42 18.65 4.75 32 19.10 4.64 19 19.56 4.53 79 19.644.52 82 20.40 4.35 24 20.69 4.29 14 21.15 4.20 28 21.49 4.13 52 22.074.02 7 22.42 3.96 16 22.86 3.89 40 23.33 3.81 16 23.81 3.73 51 24.083.69 16 24.25 3.67 22 24.57 3.62 38 25.24 3.53 22 25.40 3.50 24 25.783.45 13 26.08 3.41 48 26.45 3.37 32 26.70 3.34 22 27.12 3.29 31 27.453.25 28 27.68 3.22 13 28.13 3.17 6 29.24 3.05 28 29.56 3.02 22 29.942.98 14 30.12 2.96 22 30.39 2.94 9 31.04 2.88 32 31.29 2.86 15 31.642.83 6 32.16 2.78 8 32.69 2.74 8 32.92 2.72 21

In Table II above the value “2 θ [°]” denotes the angle of diffractionin degrees and the value “d_(hkl) [Å]” denotes the specified distancesin Å between the lattice planes.

According to the findings shown in Table II the present inventionfurther relates to the crystalline trihydrochloride trihydrate salt formofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,characterised in that in the x-ray powder diagram it has, inter alia,the characteristic values d=3.73 Å, 4.13 Å, 4.52 Å, 4.53 Å, 6.55 Å, 7.14Å, 7.73 Å, 9.01 Å and 11.21 Å (most prominent peaks in the diagram).

The material crystallizes in rod like crystals, as shown in enclosedFIG. 2.

Under standard conditions the crystalline trihydrochloride salt form ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideaccording to the invention is present in the form of an hydrate and isobtainable after drying with a changing stoichiometry (1-3 waterequivalents). The hydrate with a stoichiometry close to a trihydrateseems to be the stable hydrated form under ambient conditions and can beobtained after conditioning of the dried material. The crystal water istightly bound. Lowering the humidity down to 10% r.h. does not result ina significant weight loss.

Thus, the present invention further relates to the crystallinetrihydrochloride salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,characterised in that it is in a trihydrate form.

The thermoanalysis of the crystalline form of the crystallinetrihydrochloride salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideaccording to the invention shows a T_(fus)=245±5° C. (DSC: 10 K·min⁻¹heating rate), under decomposition, and a broad endothermic effectbetween 60-140° C., corresponding with the release of water. The DSC/TGdiagram is shown in FIG. 3. A closer look to the TG-trace shows a weightloss of approx. 7.5±0.5 wt.-% between 60 and 140° C. This weight losscan be attributed to the release of occluded crystal water.

Correlating the observed weight loss with the molecular weight of thetrihydrochloride salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamidereveals a stoichiometry of the respective hydrated form close to atrihydrate.

C₃₄H₅₀N₈O₃×3 HCl×3 H₂O M=782.26

−3 H₂O

Δm=−6.9%

C₃₄H₅₀N₈O₃×3 HCl M=728.21

At about 260° C. another strong endothermic peak can be observed in theDSC-diagram, indicating melting of the dehydrated form. The onset ofthis melting event is at about 245° C. Melting occurs, however, underdecomposition, indicated by the accompanied weight loss upon melting.When performing a TG-IR coupling experiment, it can be shown thathydrochloric acid is released upon decomposition. The respectiveanhydrous form, obtained by heating a sample up to 140° C. is notstable.

Thus, the present invention further relates to the crystallinetrihydrochloride salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,characterised by a melting point of T_(m.p.)=245±5° C. (determined byDSC; evaluation using peak-maximum; heating rate: 10° C./min).

-   -   Solid State Stability

When testing the stability of the crystalline trihydrochloride hydratedsalt form ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideunder harsh stress conditions (24 hours at 105° C., 72 hours at 70° C.and >90% rel. humid., or 24 hours in a Xenotester (k=300-800 nm, 250W·m-⁻²)), the results show that the trihydrochloride hydrated salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideis very stable in the solid state. No significant decomposition orimpurities (Σ<2.4%) can be observed under the applied stress conditions.

From the above data, it can be concluded that the hydrated form of thetrihydrochloride salt of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide of formula(I) is characterized by its high solubility in acidic and neutral mediaand its high crystallinity. The crystalline polymorph form ischaracterized as a trihydrate. The crystalline form is very stable underharsh stress conditions and is readily soluble in physiologicallyacceptable solvents. Physiologically acceptable solvents are known tothe person skilled in the art and comprise, without limitation, forexample isotonic salt or sugar containing solutions such as a 0.9% NaClsolution, a 5% glucose or mannitol solution, or a Ringer/lactatesolution.

The present invention also relates to the metabolites of the compoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamideof formula (I), to prodrugs of this compound or of these metabolitesobtained via, for example, chemical or non-chemical derivatization ofthe entire molecule or of one or more chemical groups on the molecule,to conjugates of this compound or of these metabolites with a natural orartificial polymer (for example an oligopeptide, a protein or a chemicalpolymer), and to the use thereof in a pharmaceutical composition.

“Metabolites” are intended to include compounds which are generated fromthe active parent drug according to formula (I) or other formulas orcompounds of the present invention in vivo when such parent drug isadministered to a mammalian subject.

Metabolites provide the same pharmacological effect and includecompounds of the present invention wherein for example an alkyl-aminogroup is replaced by an un-substituted amino group or the correspondingN-oxide, an ester group is replaced by the corresponding carboxylate, ora methyl group has been transformed into a hydroxymethyl or a carboxylgroup.

“Prodrugs” are intended to include any covalently bonded carriers whichrelease the active parent drug according to formula (I) or otherformulas or compounds of the present invention in vivo when such prodrugis administered to a mammalian subject. Prodrugs of a compound of thepresent invention, for example the compound of formula (I), are preparedby modifying functional groups present in the compound in such a waythat the modifications are cleaved, either in routine manipulation or invivo, to the parent compound.

“Conjugates” are intended to include any covalently bonded natural orartificial polymer (for example an oligopeptide, a protein or a chemicalpolymer) which release the active parent drug according to formula (I)or other formulas or compounds of the present invention in vivo whensuch conjugate is administered to a mammalian subject. Conjugates of acompound of the present invention, for example formula (I), are preparedby attaching functional groups present in the compound to anoligopeptide, a protein or a polymer in such a way that themodifications are cleaved in vivo by a bio-molecule, which usually isfound in the vicinity of the target, to the parent compound.

Like the dihydropteridinone derivatives mentioned in WO 2004/076454, thecompoundN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamidealso has, in particular, an inhibiting effect on specific cell cyclekinases. Thus, this compound may be used for example to treat diseasesconnected with the activity of specific cell cycle kinases andcharacterised by excessive or abnormal cell proliferation, andespecially for treating diseases connected with the activity of thepolo-like kinase PLK-1. Such diseases include, for example: viralinfections (e.g. HIV and Kaposi's sarcoma); inflammatory and autoimmunediseases (e.g. colitis, arthritis, Alzheimer's disease,glomerulonephritis and wound healing); bacterial, fungal and/orparasitic infections; leukaemias, lymphoma and solid tumours; skindiseases (e.g. psoriasis); bone diseases; cardiovascular diseases (e.g.restenosis and hypertrophy). This compound is also suitable forprotecting proliferating cells (e.g. hair, intestinal, blood andprogenitor cells) from damage to their DNA caused by radiation, UVtreatment and/or cytostatic treatment (Davis et al., 2001). It may beused for the prevention, short-term or long-term treatment of theabovementioned diseases, also in combination with other activesubstances used for the same indications, e.g. cytostatics.

Furthermore, the compounds in accordance with the present invention maybe used on their own or in conjunction with other pharmacologicallyactive substances.

Suitable preparations for the pharmaceutical compositions in accordancewith the present invention include for example tablets, capsules,suppositories, solutions,—particularly solutions for injection (s.c.,i.v., i.m.) and infusion—elixirs, emulsions or dispersible powders. Theproportion of the pharmaceutically active compound(s) should be in therange from 0.01 to 90 wt.-%, preferably 0.1 to 50 wt.-% of thecomposition as a whole, i.e. in amounts which are sufficient to achievethe dosage necessary to achieve a therapeutic effect. If necessary thedoses specified may be given several times a day.

Suitable tablets may be obtained, for example, by mixing the activesubstance(s) with known excipients, for example inert diluents such ascalcium carbonate, calcium phosphate or lactose, disintegrants such asmaize starch or alginic acid, binders such as starch or gelatine,lubricants such as magnesium stearate or talc and/or agents for delayingrelease, such as carboxymethyl cellulose, cellulose acetate phthalate,or polyvinyl acetate. The tablets may also comprise several layers.

Coated tablets may be prepared accordingly by coating cores producedanalogously to the tablets with substances normally used for tabletcoatings, for example collidone or shellac, gum arabic, talc, titaniumdioxide or sugar. To achieve delayed release or preventincompatibilities the core may also consist of a number of layers.Similarly the tablet coating may consist of a number or layers toachieve delayed release, possibly using the excipients mentioned abovefor the tablets.

Syrups or elixirs containing the active substances or combinationsthereof according to the invention may additionally contain a sweetenersuch as saccharine, cyclamate, glycerol or sugar and a flavour enhancer,e.g. a flavouring such as vanillin or orange extract. They may alsocontain suspension adjuvants or thickeners such as sodium carboxymethylcellulose, wetting agents such as, for example, condensation products offatty alcohols with ethylene oxide, or preservatives such asp-hydroxybenzoates.

Solutions for injection and infusion are prepared in the usual way, e.g.with the addition of isotonic agents, preservatives such asp-hydroxybenzoates, or stabilisers such as alkali metal salts ofethylenediamine tetraacetic acid, optionally using emulsifiers and/ordispersants, whilst if water is used as the diluent, for example,organic solvents may optionally be used as solvating agents ordissolving aids, and transferred into injection vials or ampoules orinfusion bottles.

Capsules containing one or more active substances or combinations ofactive substances may for example be prepared by mixing the activesubstances with inert carriers such as lactose or sorbitol and packingthem into gelatine capsules.

Suitable suppositories may be made for example by mixing with carriersprovided for this purpose, such as neutral fats or polyethyleneglycol orthe derivatives thereof.

Excipients which may be used include, for example, water,pharmaceutically acceptable organic solvents such as paraffins (e.g.petroleum fractions), vegetable oils (e.g. groundnut or sesame oil),mono- or polyfunctional alcohols (e.g. ethanol or glycerol), carrierssuch as e.g. natural mineral powders (e.g. kaolins, clays, talc, chalk),synthetic mineral powders (e.g. highly dispersed silicic acid andsilicates), sugars (e.g. cane sugar, lactose and glucose) emulsifiers(e.g. lignin, spent sulphite liquors, methylcellulose, starch andpolyvinylpyrrolidone) and lubricants (e.g. magnesium stearate, talc,stearic acid and sodium lauryl sulphate).

The preparations are administered by the usual methods, preferably byoral route, by injection or transdermally. For oral administration thetablets may of course contain, apart from the abovementioned carriers,additives such as sodium citrate, calcium carbonate and dicalciumphosphate together with various additives such as starch, preferablypotato starch, gelatine and the like. Moreover, lubricants such asmagnesium stearate, sodium lauryl sulphate and talc may be used at thesame time for the tabletting process. In the case of aqueous suspensionsthe active substances may be combined with various flavour enhancers orcolourings in addition to the excipients mentioned above.

For parenteral use, solutions of the active substances with suitableliquid carriers may be used.

The dosage for intravenous use is from 1-1000 mg per hour, preferablybetween 5 and 500 mg per hour.

However, it may sometimes be necessary to depart from the amountsspecified, depending on the body weight, the route of administration,the individual response to the drug, the nature of its formulation andthe time or interval over which the drug is administered. Thus, in somecases it may be sufficient to use less than the minimum dose givenabove, whereas in other cases the upper limit may have to be exceeded.When administering large amounts it may be advisable to divide them upinto a number of smaller doses spread over the day.

The following examples of formulations illustrate the present inventionwithout restricting its scope.

A) Tablets per tablet active substance 100 mg lactose 140 mg maizestarch 240 mg polyvinylpyrrolidone  15 mg magnesium stearate  5 mg 500mg

The finely ground active substance, lactose and some of the maize starchare mixed together. The mixture is screened, then moistened with asolution of polyvinylpyrrolidone in water, kneaded, wet-granulated anddried. The granules, the remaining corn starch and the magnesiumstearate are screened and mixed together. The mixture is compressed toproduce tablets of suitable shape and size.

B) Tablets per tablet active substance 80 mg lactose 55 mg maize starch190 mg  microcrystalline cellulose 35 mg polyvinylpyrrolidone 15 mgsodium-carboxymethyl starch 23 mg magnesium stearate  2 mg 400 mg 

The finely ground active substance, some of the maize starch, lactose,microcrystalline cellulose and polyvinylpyrrolidone are mixed together,the mixture is screened and worked with the remaining maize starch andwater to form a granulate which is dried and screened. The sodiumcarboxymethyl starch and the magnesium stearate are added and mixed inand the mixture is compressed to form tablets of a suitable size.

C) Ampoule solution active substance 50 mg sodium chloride 900 mg sodiumhydroxide up to pH 4.0 water for inj. q.s.p. 100 ml

The active substance is dissolved in water at its own pH and sodiumchloride is added to make the solution isotonic. The pH is then adjustedto 4.0 by addition of sodium hydroxide 1 N. The resulting solution isfiltered to remove pyrogens and the filtrate is transferred underaseptic conditions into ampoules which are then sterilised andheat-sealed. The ampoules may contain 5 mg, 25 mg or 50 mg of activesubstance.

1. The trihydrochloride salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.2. A crystalline, hydrated form of the trihydrochloride salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide.3. A crystalline, trihydrated form of the trihydrochloride salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide, inaccordance with claim
 2. 4. The crystalline form of the trihydrate ofthe trihydrochloride trihydrate salt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,in accordance with claim 3, having the following characteristic valuesd: 3.73 Å, 4.13 Å, 4.52 Å, 4.53 Å, 6.55 Å, 7.14 Å, 7.73 Å, 9.01 Å and11.21 Å.
 5. A pharmaceutical composition comprising the trihydrochloridesalt ofN-[trans-4-[4-(cyclopropylmethyl)-1-piperazinyl]cyclohexyl]-4-[[(7R)-7-ethyl-5,6,7,8-tetrahydro-5-methyl-8-(1-methylethyl)-6-oxo-2-pteridinyl]amino]-3-methoxy-benzamide,or the crystalline form thereof in accordance with claim 2, 3 or 4,together with a pharmaceutically acceptable carrier or diluent.